LOCUS       sequence-80206-e         911 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 911)
  AUTHORS   Stanton BC, Nielsen AA, Tamsir A, Clancy K, Peterson T, Voigt CA
  TITLE     Genomic mining of prokaryotic repressors for orthogonal logic gates.
  JOURNAL   Nat Chem Biol. 2013 Dec 8. doi: 10.1038/nchembio.1411.
  PUBMED    24316737
REFERENCE   2  (bases 1 to 911)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem 
            Biol. 2013 Dec 8. doi: 10.1038/nchembio.1411."
COMMENT     Sequence Label: pET21a-PqrA
FEATURES             Location/Qualifiers
     source          1..911
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     60..79
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        60..78
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    79..103
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             118..140
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             159..176
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             186..209
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             792..809
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(862..880)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt
       61 aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt
      121 gtttaacttt aagaaggaga tataccatgg gcagcagcca tcatcatcat catcacagca
      181 gcggcctgga agttctgttc caggggcccc atatggcagt tgatcgtgaa caggttctgc
      241 gtagcgcagc aaccctgctg acccgtaaaa gcaccgcaac catggatgaa gttgcacgtg
      301 cagcaggtct gagccgtgca accctgcatc gtcattttgc aggtcgtgat gcactggttc
      361 gtgcactgga aagcctgggt attgcagaat gtgaagcagc actggcagca gcacgtaccg
      421 atgaaggtcc ggcagcagat gcagttcgtc gtctggttcg tgaaatggaa ccgagcgcag
      481 cactgctggc atttctgtat accgaaaatc agctgtttga aggcgaagaa cagaatgaag
      541 gttgggcacg tattgatacc ggtctgaccg aactgtttcg tcgtggtcaa gaaagcggtg
      601 aatttcgtat tgatctgaca ccggcatggc tgaccgaagc actgtatggt ctgctggcaa
      661 gcggtgcatg ggcagttacc gaaggtcgtg ttgcacgtaa tgattttacc cacatgattg
      721 ttgaactgct gctgggtggt gcactgcgtc gtgaagaacc gtaaggatcc aagcttgcgg
      781 ccgcactcga gcaccaccac caccaccact gagatccggc tgctaacaaa gcccgaaagg
      841 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
      901 aacgggtctt g
//
